PDF(Pubmed)
Abstract：
Toscana virus (TOSV), a sandfly-borne virus, is an important etiological agent in human acute meningitis and meningoencephalitis in the Mediterranean area during the summer. However, the actual number of TOSV infections is underestimated. Laboratory confirmation is necessary because TOSV infection has overlapping clinical features with other neuro-invasive viral infections. Nowadays, the reference test for direct diagnosis in the acute phase of TOSV infection is the PCR based method for detecting TOSV in cerebrospinal fluid and/or plasma, serum, or blood. Although poorly employed, urine is another helpful biological matrix for TOSV detection. Urine is a matrix rich in PCR inhibitors that affect PCR efficiency; consequently, false negatives could be generated. To investigate the potential effect of urine PCR inhibitors on TOSV detection, we compared undiluted and diluted urine using 10-fold series of spiked TOSV. The results showed a significant improvement in TOSV detection performance in diluted urine (1 TCID50 vs. 1 × 104 TCID50 limit of detection and 101.35% vs. 129.62% efficiency, respectively, in diluted and undiluted urine). In conclusion, our data provide preliminary important insights into the use of diluted urine to limit the impact of the inhibitory effects of urine on the detection of TOSV in RT-PCR-based approaches.
摘要：
托斯卡纳病毒（TOSV），沙蝇传播的病毒，是夏季地中海地区人类急性脑膜炎和脑膜脑炎的重要病因。然而,TOSV感染的实际数量被低估了.实验室确认是必要的，因为TOSV感染与其他神经侵袭性病毒感染具有重叠的临床特征。如今,在TOSV感染急性期直接诊断的参考测试是基于PCR的方法，用于检测脑脊液和/或血浆中的TOSV，血清,或者血.虽然就业差，尿液是另一种有助于TOSV检测的生物基质。尿液是富含影响PCR效率的PCR抑制剂的基质；因此，可能会产生假阴性。探讨尿液PCR抑制剂对TOSV检测的潜在影响，我们使用10倍系列加标TOSV比较了未稀释和稀释的尿液。结果表明，稀释尿液中的TOSV检测性能显着改善（1TCID50与1×104TCID50检测限和101.35%vs.129.62%效率,分别,在稀释和未稀释的尿液中）。总之,我们的数据为使用稀释的尿液限制尿液对基于RT-PCR的TOSV检测的抑制作用的影响提供了初步的重要见解.
